Spinach hybrid 51-330

ABSTRACT

The present invention relates to the development of a spinach plant exhibiting a combination of traits which may include resistance against downy mildew ( Peronospora farinosa  f. sp. Spinaciae) strains Pf1-Pf13, and arrow-shaped prime market stage leaves, methods and compositions relating to plants, progeny, seeds and derivatives thereof and plants, progeny, seeds and derivatives thereof having all the morphological, physiological and/or genetic characteristics thereof.

RELATED APPLICATIONS AND INCORPORATION BY REFERENCE

This application claims priority to U.S. provisional patent applicationSer. No. 61/528,947 filed Aug. 30, 2011.

The foregoing applications, and all documents cited therein or duringtheir prosecution (“appln cited documents”) and all documents cited orreferenced in the appln cited documents, and all documents cited orreferenced herein (“herein cited documents”), and all documents cited orreferenced in herein cited documents, together with any manufacturer'sinstructions, descriptions, product specifications, and product sheetsfor any products mentioned herein or in any document incorporated byreference herein, are hereby incorporated herein by reference, and maybe employed in the practice of the invention. More specifically, allreferenced documents are incorporated by reference to the same extent asif each individual document was specifically and individually indicatedto be incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to the development of a spinach plantexhibiting a combination of traits which may include resistance againstdowny mildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13,and arrow-shaped prime market stage leaves.

BACKGROUND OF THE INVENTION

Spinach (Spinacia oleracea) is a flowering vegetable plant in the familyof Amaranthaceae. It is native to southwestern and central Asia, butnowadays is being cultivated worldwide, mostly in temperate regions. Theconsumable parts of spinach are the leaves. These are produced duringthe first stage of the life cycle of a spinach plant, during which theplant forms a leaf rosette. The second stage is the flowering stage orbolting stage. Bolting is the growth of an elongated stalk with flowersgrown from within the main stem of a plant. During the bolting stage itis not possible anymore to harvest any marketable product of the plant.For the start of bolting different regional varieties appear to havedifferent starting moments. The variety “Dixie Market” expresses anearly start of bolting; the variety “Long Standing Bloomsdale” shows amedium start of bolting whereas the variety “Norgreen” initiates a latestart of bolting.

The leaves of a spinach plant are usually sold loose, bunched, inprepackaged bags, canned, or frozen. There are three basic types ofspinach, namely savoy, semi-savoy and smooth. Savoy has dark green,crinkly and curly leaves. Flat or smooth leaf spinach has broad smoothleaves. Semi-savoy is a hybrid variety with slightly crinkled leaves.

Downy mildew is probably the most widespread and potentially destructiveglobal disease of spinach. The causal agent of downy mildew disease onvarious plants of Chenopodiaceae, including spinach, is regarded as asingle species, Peronospora farinosa. Some of the forms on importantcrop plants have been given names as formae speciales, so f. sp.spinaciae on spinach. Downy mildew affects the harvested part (leaves)of spinach (Spinacia oleracea). Yellow lesions appear on the olderleaves. Although some fungicide treatments are effective, they arecostly and cause ecological pollution. There is a need for resistantspinach cultivars. There are currently 13 recognised described P.farinosa races (races Pf1 to Pf13) of P. farinosa f. sp. Spinaciae and anumber of isolates not officially recognized yet. Although the pathogenwas first reported early in the nineteenth century, only three races ofthe pathogen had been identified before 1990. More than ten new races ofthe pathogen were identified between 1990 and 2010, and some of thenewer races had overcome all known genetic resistance. The rapidascendance of new races is likely to be a result of intensification andscaling-up in spinach production during the past decade.

Citation or identification of any document in this application is not anadmission that such document is available as prior art to the presentinvention.

SUMMARY OF THE INVENTION

In one embodiment, the invention provides a spinach plant exhibiting acombination of traits which may include resistance against downy mildew(Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, andarrow-shaped prime market stage leaves, representative seed of whichhaving been deposited under NCIMB Accession NCIMB 41821.

In one embodiment, the invention provides a spinach plant exhibiting acombination of traits which may include resistance against downy mildew(Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, arrow-shapedprime market stage leaves, and prime market stage leaves with a slightlycurled up margin, representative seed of which having been depositedunder NCIMB Accession NCIMB 41821.

In one embodiment, the invention provides a spinach plant exhibiting acombination of traits which may include resistance against downy mildew(Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, arrow-shapedprime market stage leaves, prime market stage leaves with a slightlycurled up margin, and medium start of bolting, representative seed ofwhich having been deposited under NCIMB Accession NCIMB 41821.

In one embodiment, the invention provides a spinach plant designated51-330, representative seed of which having been deposited under NCIMBAccession NCIMB 41821.

In one embodiment there is a plant grown from seeds, representative seedof which have been deposited under NCIMB Accession NCIMB 41821.

As used herein, a plant of the invention and/or a spinach plant of theinvention may encompass a spinach plant having, expressing and showing anew resistance against downy mildew (Peronospora farinosa f. sp.Spinaciae) strains Pf1-Pf13, arrow-shaped prime market stage leaves,prime market stage leaves with a slightly curled up margin and/or mediumstart of bolting. A plant of the invention and/or a spinach plant of theinvention may be designated 51-330 or have all the morphological,physiological and/or genetic characteristics of a hybrid spinach variety51-330, representative seed of which having been deposited under NCIMBAccession NCIMB 41821.

In one embodiment the present invention relates to parts of a spinachplant of the invention, a spinach plant exhibiting a combination oftraits which may include resistance against downy mildew (Peronosporafarinosa f. sp. Spinaciae) strains Pf1-Pf13, and arrow-shaped primemarket stage leaves, wherein the plant parts may be involved in sexualreproduction, which may include, without limitation, microspores,pollen, ovaries, ovules, embryo sacs or egg cells

In addition, the invention relates to parts of the plant that aresuitable for vegetative reproduction, which may include, withoutlimitation, cuttings, roots, stems, cells, and protoplasts of thespinach plants of the invention.

In one embodiment the invention relates to a tissue culture ofregenerable cells of a plant of the invention. Such a tissue culture maybe derived, without limitation, from leaves, microspores, pollen,ovaries, ovules, embryo sacs or egg cells, cotyledon, hypocotyls,meristematic cells, roots, root tips, anthers, flowers, seeds and stems.The plants of the invention from which such parts may come from, includethose of which representative seed been deposited under NCIMB AccessionNCIMB 41821.

In one embodiment, there is provided progeny of a spinach plant whichmay exhibit a combination of traits including resistance against downymildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, andarrow-shaped prime market stage leaves, produced by sexual or vegetativereproduction, grown from seeds, regenerated from the above-describedplant parts, or regenerated from the above-described tissue culture ofthe hybrid spinach variety or a progeny plant thereof, representativeseed of which have been deposited under NCIMB Accession NCIMB 41821.

In another embodiment, there is provided progeny of a spinach plantexhibiting a combination of traits which may include resistance againstdowny mildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13,and arrow-shaped prime market stage leaves, produced by sexual orvegetative reproduction, grown from seeds, regenerated from theabove-described plant parts, or regenerated from the above-describedtissue culture of the spinach plant of the invention or a progeny plantthereof, in which the regenerated plant may exhibit a combination oftraits, including resistance against downy mildew (Peronospora farinosaf. sp. Spinaciae) strains Pf1-Pf13, and arrow-shaped prime market stageleaves, representative seed of which having been deposited under NCIMBAccession NCIMB 41821.

Progeny of a spinach plant exhibiting a combination of traits includingresistance against downy mildew (Peronospora farinosa f. sp. Spinaciae)strains Pf1-Pf13, and arrow-shaped prime market stage leaves, may bemodified in one or more other characteristics, in which the modificationis a result of, for example and without limitation, mutagenesis ortransformation with a transgene.

In one embodiment, there is provided progeny of a spinach plant whichmay exhibit a combination of traits including resistance against downymildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, andarrow-shaped prime market stage leaves, produced by sexual or vegetativereproduction, grown from seeds, regenerated from the above-describedplant parts, or regenerated from the above-described tissue culture ofhybrid spinach variety or a progeny plant thereof, in which theregenerated plant may exhibit a combination of traits includingresistance against downy mildew (Peronospora farinosa f. sp. Spinaciae)strains Pf1-Pf13, and arrow-shaped prime market stage leaves,representative seed of which having been deposited under NCIMB AccessionNCIMB 41821, and is modified in one or more other traits.

In another embodiment the invention relates to a method of producing aninbred spinach plant derived from a plant of the invention of whichrepresentative seed has been deposited under NCIMB Accession NCIMB41821, which may comprise steps of: a) preparing a progeny plant derivedfrom a spinach plant which may exhibit a combination of traits includingresistance against downy mildew (Peronospora farinosa f. sp. Spinaciae)strains Pf1-Pf13, and arrow-shaped prime market stage leaves by crossingthe spinach plant of the invention, being a spinach plant which mayexhibit a combination of traits including resistance against downymildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, andarrow-shaped prime market stage leaves, representative seed of whichhaving been deposited under NCIMB Accession NCIMB 41821 with a secondspinach plant; b) crossing the progeny plant with itself or a secondspinach plant to produce a seed of a progeny plant of a subsequentgeneration; c) growing a progeny plant of a subsequent generation fromsaid seed and crossing the progeny plant of a subsequent generation withitself or a second spinach plant; and d) repeating step b) or c) for atleast 1 more generation to produce an inbred spinach plant derived froma spinach plant which may exhibit a combination of traits includingresistance against downy mildew (Peronospora farinosa f. sp. Spinaciae)strains Pf1-Pf13, and arrow-shaped prime market stage leaves.

It is noted that in this disclosure and particularly in the claims,terms such as “comprises”, “comprised”, and “comprising” and the like(e.g., “includes”, “included”, “including”, “contains”, ‘contained”,“containing”, “has”, “had”, “having”, etc.) can have the meaningascribed to them in US Patent law, i.e., they are open ended terms. Forexample, any method that “comprises,” “has” or “includes” one or moresteps is not limited to possessing only those one or more steps and alsocovers other unlisted steps. Similarly, any plant that “comprises,”“has” or “includes” one or more traits is not limited to possessing onlythose one or more traits and covers other unlisted traits. Similarly,the terms “consists essentially of” and “consisting essentially of” havethe meaning ascribed to them in US Patent law, e.g., they allow forelements not explicitly recited, but exclude elements that are found inthe prior art or that affect a basic or novel characteristic of theinvention. See also MPEP §2111.03. In addition, the term “about” is usedto indicate that a value includes the standard deviation of error forthe device or method being employed to determine the value.

These and other embodiments are disclosed or are obvious from andencompassed by the following Detailed Description.

Deposit

The Deposit with NCIMB Ltd, Ferguson Building, Craibstone Estate,Bucksburn, Aberdeen AB21 9YA, UK, on Mar. 17, 2011, under depositaccession number NCIMB 41821 was made pursuant to the terms of theBudapest Treaty. Upon issuance of a patent, all restrictions upon thedeposit will be removed, and the deposit is intended to meet therequirements of 37 CFR §1.801-1.809. The deposit will be maintained inthe depository for a period of 30 years, or 5 years after the lastrequest, or for the effective life of the patent, whichever is longer,and will be replaced if necessary during that period.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description, including the examples, given by wayof example, but not intended to limit the invention solely to thespecific embodiments described, may be best understood in conjunctionwith the accompanying drawings.

FIG. 1 shows the leaf, the leaf base and leaf tip shapes of firstfoliage leaves.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of spinach plants a new combination of traitswhich may include resistance against downy mildew (Peronospora farinosaf. sp. Spinaciae) strains Pf1-Pf13, and arrow-shaped prime market stageleaves. A spinach plant with this new characteristics will be hereinreferred to as hybrid spinach variety 51-330. Hybrid spinach variety51-330 is a commercial spinach hybrid distinct from other such hybrids.

The parents of hybrid spinach variety 51-330 were developed as follows:The mother is a spinach line made from pedigree selection fromLombardia. In total 5 selection, crossing, mass generation and selfingcycles were performed, selecting for uniformity. The father line of51-330 is an inbred line selection from 06.98952, obtained by 5succeeding selection and inbreeding cycles.

Crossing the described mother and father inbred spinach lines with oneanother will yield uniform F1 hybrid progeny plants. Table 1 shows thepedigree scheme of the mother line, table 2 shows the pedigree scheme ofthe father line of hybrid spinach line 51-330.

The F1 may be self-pollinated to produce a segregating F2 generation.Individual plants may then be selected which represent the desiredphenotype in each generation (F3, F4, F5, etc.) until the traits arehomozygous or fixed within a breeding population.

TABLE 1 Breeding history of the mother line of 51-330 (M = Massselection, S = Selfing) Year Year 1 F1 Pedigree selection from LombardiaYear 2 M1 F1 generation grown Year 3 S1 M1 F1 generation grown Year 4 M1S1 M1 F1 generation grown (in mass) Year 5 BC1 M1 S1 M1 F1 generationgrown

TABLE 2 Breeding history of the father line of 51-330 Year Year 1 F1Pedigree selection from 06.98952 Year 2 S1 F1 generation grown Year 3 S2F1 generation grown Year 4 S3 F1 generation grown Year 5 S4 F1generation grown

In one embodiment, a plant of the invention has all the morphologicaland physiological characteristics of hybrid spinach variety 51-330.

The physiological and morphological differences with regard to the primemarket stage leaf characteristics and bolting between hybrid spinachvariety 51-330 and closest known varieties “Swan” and “Whale” aresummarized in table 3.

Embodiments of the invention advantageously have one or more, and mostadvantageously all, of these characteristics.

TABLE 3 Physiological and morphological characteristics of hybridspinach variety 51-330 with closest known varieties “Swan” and “Whale”Cultivar Line 51-330 Swan Whale For patent or patent comparisoncomparison comparison Characteristics Species Spinacia oleracea L.Spinacia oleracea L. Spinacia oleracea L. Ploidy Diploid Diploid DiploidMaturity Growth Rate Medium (Long Standing Medium (Long Standing Slow(Nogreen) Bloomsdale) Bloomsdale) Plant (prime market stage) HabitSemi-erect (Long Semi-erect (Long Semi-erect (Long Standing Bloomsdale)Standing Bloomsdale) Standing Bloomsdale) Size Medium Large (GiantNobel) Medium Spread (cm) 44 58 52 Height (cm) 29 28 28 SeedlingCotyledon Width (mm) 6 7 7 Length (mm) 68 47 68 Tip Pointed RoundedPointed Color Color Chart Name RHS CC RHS CC RHS CC Color Chart Value146A 146A 146B Leaf (First Foliage Leaves) Shape Elliptic CircularElliptic Base Straight Straight Straight Tip Round-pointed RoundRound-pointed Margin Slightly Curled Slightly Curled Curled under UpperSurface Color Color Chart Name RHS CC RHS CC RHS CC Color Chart Value146A 146A 146A Lower surface Color Lighter Lighter Lighter (comparedwith upper) Color Chart Name RHS CC RHS CC RHS CC Color Chart Value 146B146B 146B Leaf (Prime Market Stage) Surface Smooth to Semi SavoySemi-savoy Smooth to Semi Savoy Shape Arrow-shaped Five-sided Five-sidedBase Straight Straight V-shape Tip Pointed Round-pointed Round-pointedMargin Slightly curled up Slightly Curled Curled under Upper SurfaceColor Color Chart Name RHS CC RHS CC RHS CC Color Chart Value 146A 146A146A Lower surface Color Lighter Lighter Lighter (compared with upper)Color Chart Name RHS CC RHS CC RHS CC Color Chart Value 144A 144A 146BLuster Dull Glossy Glossy Blade Size Medium (Virginia Large (GiantNobel) Medium (Virginia Savoy) Savoy) Blade Lobing Lobed Lobed LobedPetiole Color Light Green White White to Light Green Color Chart NameRHS CC RHS CC RHS CC Color Chart Value 144B 144A 144A Petiole Length to20 21 22 the Blade (cm) Petiole Diameter 5 10 8 (mm) Seed StalkDevelopment Start of bolting Medium (Long Standing Medium (Long StandingMedium (Long Standing (10% of the plants) Bloomsdale) Bloomsdale)Bloomsdale)

In an embodiment, the invention relates to spinach plants that have allthe morphological and physiological characteristics of the invention andhave acquired said characteristics by introduction of the geneticinformation that is responsible for the characteristics from a suitablesource, either by conventional breeding, or genetic modification, inparticular by cisgenesis or transgenesis.

Cisgenesis is genetic modification of plants with a natural gene, codingfor an (agricultural) trait, from the crop plant itself or from asexually compatible donor plant. Transgenesis is genetic modification ofa plant with a gene from a non-crossable species or a synthetic gene.

Just as useful traits that may be introduced by backcrossing, usefultraits may be introduced directly into the plant of the invention, beinga plant of hybrid spinach line 51-330, by genetic transformationtechniques; and, such plants of the invention that have additionalgenetic information introduced into the genome or that expressadditional traits by having the DNA coding there for introduced into thegenome via transformation techniques, are within the ambit of theinvention, as well as uses of such plants, and the making of suchplants.

Genetic transformation may therefore be used to insert a selectedtransgene into the plant of the invention, being a plant of hybridspinach line 51-330 or may, alternatively, be used for the preparationof transgenes which may be introduced by backcrossing. Methods for thetransformation of plants, including spinach, are well known to those ofskill in the art.

Vectors used for the transformation of spinach cells are not limited solong as the vector may express an inserted DNA in the cells. Forexample, vectors which may comprise promoters for constitutive geneexpression in spinach cells (e.g., cauliflower mosaic virus 35Spromoter) and promoters inducible by exogenous stimuli may be used.Examples of suitable vectors include pBI binary vector. The “spinachcell” into which the vector is to be introduced includes various formsof spinach cells, such as cultured cell suspensions, protoplasts, leafsections, and callus. A vector may be introduced into spinach cells byknown methods, such as the polyethylene glycol method, polycationmethod, electroporation, Agrobacterium-mediated transfer, particlebombardment and direct DNA uptake by protoplasts. To effecttransformation by electroporation, one may employ either friabletissues, such as a suspension culture of cells or embryogenic callus oralternatively one may transform immature embryos or other organizedtissue directly. In this technique, one would partially degrade the cellwalls of the chosen cells by exposing them to pectin-degrading enzymes(pectolyases) or mechanically wound tissues in a controlled manner.

A particularly efficient method for delivering transforming DNA segmentsto plant cells is microprojectile bombardment. In this method, particlesare coated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate. An illustrativeembodiment of a method for delivering DNA into plant cells byacceleration is the Biolistics Particle Delivery System, which may beused to propel particles coated with DNA or cells through a screen, suchas a stainless steel or Nytex screen, onto a surface covered with targetspinach cells. The screen disperses the particles so that they are notdelivered to the recipient cells in large aggregates. It is believedthat a screen intervening between the projectile apparatus and the cellsto be bombarded reduces the size of projectiles aggregate and maycontribute to a higher frequency of transformation by reducing thedamage inflicted on the recipient cells by projectiles that are toolarge. Microprojectile bombardment techniques are widely applicable, andmay be used to transform virtually any plant species, including a plantof hybrid spinach line 51-330.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA may be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast.Agrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations.Moreover, advances in vectors for Agrobacterium-mediated gene transferhave improved the arrangement of genes and restriction sites in thevectors to facilitate the construction of vectors capable of expressingvarious polypeptide coding genes. The vectors have convenientmulti-linker regions flanked by a promoter and a polyadenylation sitefor direct expression of inserted polypeptide coding genes.Additionally, Agrobacterium containing both armed and disarmed Ti genesmay be used for transformation. In those plant strains whereAgrobacterium-mediated transformation is efficient, it is the method ofchoice because of the facile and defined nature of the gene locustransfer. The use of Agrobacterium-mediated plant integrating vectors tointroduce DNA into plant cells, including spinach plant cells, is wellknown in the art (See, e.g., U.S. Pat. Nos. 7,250,560 and 5,563,055).

Transformation of plant protoplasts also may be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments.

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for spinach plant geneexpression include, but are not limited to, the cauliflower mosaic virus(CaMV) P-35S promoter, a tandemly duplicated version of the CaMV 35Spromoter, the enhanced 35S promoter (P-e35S), the nopaline synthasepromoter, the octopine synthase promoter, the figwort mosaic virus(P-FMV) promoter (see U.S. Pat. No. 5,378,619), an enhanced version ofthe FMV promoter (P-eFMV) where the promoter sequence of P-FMV isduplicated in tandem, the cauliflower mosaic virus 19S promoter, asugarcane bacilliform virus promoter, a commelina yellow mottle viruspromoter, the promoter for the thylakoid membrane proteins from spinach(psaD, psaF, psaE, PC, FNR, atpC, atpD, cab, rbcS) (see U.S. Pat. No.7,161,061), the CAB-1 promoter from spinach (see U.S. Pat. No.7,663,027), the promoter from maize prolamin seed storage protein (seeU.S. Pat. No. 7,119,255), and other plant DNA virus promoters known toexpress in plant cells. A variety of plant gene promoters that areregulated in response to environmental, hormonal, chemical, and/ordevelopmental signals may be used for expression of an operably linkedgene in plant cells, including promoters regulated by (1) heat, (2)light (e.g., pea rbcS-3A promoter, maize rbcS promoter, or chlorophylla/b-binding protein promoter), (3) hormones, such as abscisic acid, (4)wounding (e.g., wunl, or (5) chemicals such as methyl jasmonate,salicylic acid, or Safener. It may also be advantageous to employorgan-specific promoters.

Exemplary nucleic acids which may be introduced to the spinach plant ofthis invention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin spinach species, but are incorporated into recipient cells by geneticengineering methods rather than classical reproduction or breedingtechniques. However, the term “exogenous” is also intended to refer togenes that are not normally present in the cell being transformed, orperhaps simply not present in the form, structure, etc., as found in thetransforming DNA segment or gene, or genes which are normally presentand that one desires to express in a manner that differs from thenatural expression pattern, e.g., to over-express. Thus, the term“exogenous” gene or DNA is intended to refer to any gene or DNA segmentthat is introduced into a recipient cell, regardless of whether asimilar gene may already be present in such a cell. The type of DNAincluded in the exogenous DNA may include DNA which is already presentin the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a plant of the invention. Non-limitingexamples of particular genes and corresponding phenotypes one may chooseto introduce into a spinach plant include one or more genes for insecttolerance, pest tolerance such as genes for fungal disease control,herbicide tolerance, and genes for quality improvements such as yield,nutritional enhancements, environmental or stress tolerances, or anydesirable changes in plant physiology, growth, development, morphologyor plant product(s).

Alternatively, the DNA coding sequences may affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms. The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product. Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention. (See also U.S. Pat. No.7,576,262, “Modified gene-silencing RNA and uses thereof.”

U.S. Pat. Nos. 7,230,158, 7,122,720, 7,081,363, 6,734,341, 6,503,732,6,392,121, 6,087,560, 5,981,181, 5,977,060, 5,608,146, 5,516,667, eachof which, and all documents cited therein are hereby incorporated hereinby reference, consistent with the above INCORPORATION BY REFERENCEsection, are additionally cited as examples of U.S. Patents that mayconcern transformed spinach and/or methods of transforming spinach orspinach plant cells, and techniques from these US Patents, as well aspromoters, vectors, etc., may be employed in the practice of thisinvention to introduce exogenous nucleic acid sequence(s) into a plantof hybrid spinach line 51-330 (or cells thereof), and exemplify someexogenous nucleic acid sequence(s) which may be introduced into a plantof hybrid spinach line 51-330 (or cells thereof) of the invention, aswell as techniques, promoters, vectors etc., to thereby obtain furtherplants of hybrid spinach line 51-330, plant parts and cells, seeds,other propagation material harvestable parts of these plants, etc. ofthe invention, e.g. tissue culture, including a cell or protoplast, suchas an embryo, meristem, cotyledon, pollen, leaf, anther, root, root tip,pistil, flower, seed or stalk.

The invention further relates to propagation material for producingplants of the invention. Such propagation material may comprise interalia seeds of the claimed plant and parts of the plant that are suitablefor sexual reproduction. Such parts are for example selected from thegroup consisting of seeds, microspores, pollen, ovaries, ovules, embryosacs and egg cells. In addition, the invention relates to propagationmaterial which may comprise parts of the plant that are suitable forvegetative reproduction, for example cuttings, roots, stems, cells,protoplasts.

According to a further aspect thereof the propagation material of theinvention may comprise a tissue culture of the claimed plant. The tissueculture may comprise regenerable cells. Such tissue culture may bederived from leaves, pollen, embryos, cotyledon, hypocotyls,meristematic cells, roots, root tips, anthers, flowers, seeds and stems.(See generally U.S. Pat. No. 7,041,876 on spinach being recognized as aplant that may be regenerated from cultured cells or tissue).

Also, the invention comprehends methods for producing a seed of a“51-330”-derived spinach plant which may comprise (a) crossing a plantof hybrid spinach variety 51-330, representative seed of which havingbeen deposited under NCIMB Accession NCIMB 41821, with a second spinachplant, and (b) whereby seed of a “51-330”-derived spinach plant form(e.g., by allowing the plant from the cross to grow to producing seed).Such a method may further comprise (c) crossing a plant grown from“51-330”-derived spinach seed with itself or with a second spinach plantto yield additional “51-330”-derived spinach seed, (d) growing theadditional “51-330”-derived spinach seed of step (c) to yield additional“51-330”-derived spinach plants, and (e) repeating the crossing andgrowing of steps (c) and (d) to generate further “51-330”-derivedspinach plants.

The invention further involves a method of determining the genotype of aplant of hybrid spinach variety 51-330, representative seed of which hasbeen deposited under NCIMB Accession NCIMB 41821, or a first generationprogeny thereof, which may comprise obtaining a sample of nucleic acidsfrom said plant and detecting in said nucleic acids a plurality ofpolymorphisms. This method may additionally comprise the step of storingthe results of detecting the plurality of polymorphisms on a computerreadable medium and/or transmitting the results of detecting theplurality of polymorphisms, e.g., by telephony or by means of computer(e.g., via email). The plurality of polymorphisms are indicative ofand/or give rise to the expression of the morphological andphysiological characteristics of hybrid spinach variety 51-330.

The polymorphisms of this invention may be provided in a variety ofmediums to facilitate use, e.g. a database or computer readable medium,which may also contain descriptive annotations in a form that allows askilled artisan to examine or query the polymorphisms and obtain usefulinformation.

As used herein “database” refers to any representation of retrievablecollected data including computer files such as text files, databasefiles, spreadsheet files and image files, printed tabulations andgraphical representations and combinations of digital and image datacollections. In a preferred aspect of the invention, “database” refersto a memory system that may store computer searchable information.

As used herein, “computer readable media” refers to any medium that maybe read and accessed directly by a computer. Such media include, but arenot limited to: magnetic storage media, such as floppy discs, hard disc,storage medium and magnetic tape; optical storage media such as CD-ROM;electrical storage media such as RAM, DRAM, SRAM, SDRAM, ROM; and PROMs(EPROM, EEPROM, Flash EPROM), and hybrids of these categories such asmagnetic/optical storage media. A skilled artisan may readily appreciatehow any of the presently known computer readable mediums may be used tocreate a manufacture which may comprise computer readable medium havingrecorded thereon a polymorphism of the present invention.

As used herein, “recorded” refers to the result of a process for storinginformation in a retrievable database or computer readable medium. Forinstance, a skilled artisan may readily adopt any of the presently knownmethods for recording information on computer readable medium togenerate media which may comprise the polymorphisms of the presentinvention. A variety of data storage structures are available to askilled artisan for creating a computer readable medium where the choiceof the data storage structure will generally be based on the meanschosen to access the stored information. In addition, a variety of dataprocessor programs and formats may be used to store the polymorphisms ofthe present invention on computer readable medium.

The present invention further provides systems, particularlycomputer-based systems, which contain the polymorphisms describedherein. Such systems are designed to identify the polymorphisms of thisinvention. As used herein, “a computer-based system” refers to thehardware, software and memory used to analyze the polymorphisms. Askilled artisan may readily appreciate that any one of the currentlyavailable computer-based system are suitable for use in the presentinvention.

Spinach leaves are sold in packaged form, including without limitationas pre-packaged spinach salad or as canned spinach or as frozen spinach.Mention is made of U.S. Pat. No. 5,523,136, incorporated herein byreference consistent with the above INCORPORATION BY REFERENCE section,which provides packaging film, and packages from such packaging film,including such packaging containing leafy produce, and methods formaking and using such packaging film and packages, which are suitablefor use with the spinach leaves of the invention. Thus, the inventioncomprehends the use of and methods for making and using the leaves ofthe spinach of the invention, as well as leaves of spinach derived fromthe invention. The invention further relates to a container which maycomprise one or more plants of the invention, or one or spinach plantsderived from a plant of the invention, in a growth substrate for harvestof leaves from the plant in a domestic environment. This way theconsumer may pick very fresh leaves for use in salads. More generally,the invention includes one or more plants of the invention or one ormore plants derived from spinach of the invention, wherein the plant isin a ready-to-harvest condition, including with the consumer picking hisown, and further including a container which may comprise one or more ofthese plants.

The invention is further described by the following numbered paragraphs:

-   1. A spinach plant exhibiting a combination of traits including    resistance against downy mildew (Peronospora farinosa f. sp.    Spinaciae) strains Pf1-Pf13, and arrow-shaped prime market stage    leaves, representative seed of which having been deposited under    NCIMB Accession NCIMB 41821.-   2. A spinach plant exhibiting a combination of traits including    resistance against downy mildew (Peronospora farinosa f. sp.    Spinaciae) strains Pf1-Pf13, arrow-shaped prime market stage leaves,    and prime market stage leaves with a slightly curled up margin,    representative seed of which having been deposited under NCIMB    Accession NCIMB 41821.-   3. A spinach plant exhibiting a combination of traits including    resistance against downy mildew (Peronospora farinosa f. sp.    Spinaciae) strains Pf1-Pf13, arrow-shaped prime market stage leaves,    prime market stage leaves with a slightly curled up margin, and    medium start of bolting, representative seed of which having been    deposited under NCIMB Accession NCIMB 41821.-   4. A hybrid spinach variety designated 51-330, representative seed    of which have deposited under NCIMB Accession NCIMB 41821.-   5. A spinach plant having all the morphological and physiological    characteristics of a plant as paragraph 4, representative seed of    which have deposited under NCIMB Accession NCIMB 41821.-   6. Seed of the plant of paragraph 1 or paragraph 2 or paragraph 3 or    paragraph 4 or paragraph 5.-   7. Parts of the plant of paragraph 1 or paragraph 2 or paragraph 3    or paragraph 4 or paragraph 5, wherein said parts of the plant are    suitable for sexual reproduction.-   8. Parts of the plant as paragraph 7, said parts selected from the    group consisting of microspores, pollen, ovaries, ovules, embryo    sacs and egg cells.-   9. Parts of the plant of paragraph 1 or paragraph 2 or paragraph 3    or paragraph 4 or paragraph 5, wherein said parts of the plant are    suitable for vegetative reproduction.-   10. Parts as paragraph 9, said parts selected from the group    consisting of cuttings, roots, stems, cells and protoplasts.-   11. A tissue culture of regenerable cells from the spinach plant of    paragraph 1 or paragraph 2 or paragraph 3 or paragraph 4 or    paragraph 5.-   12. A tissue culture as paragraph 11, wherein said cells or    protoplasts of the tissue culture which are derived from a tissue    selected from the group consisting of leaves, pollen, embryos,    cotyledon, hypocotyls, meristematic cells, roots, root tips,    anthers, flowers, seeds and stems.-   13. Progeny of a spinach plant of paragraph for paragraph 2 or    paragraph 3 or paragraph 4 or paragraph 5.-   14. Progeny as paragraph 13, wherein said progeny is produced by    sexual or vegetative reproduction of said spinach plant.-   15. Progeny of a spinach plant of paragraph 1 or paragraph 2 or    paragraph 3 or paragraph 4 or paragraph 5, wherein the plant    exhibits a combination of traits including resistance against downy    mildew (Peronospora farinosa f. sp. Spinaciae) strains Pf1-Pf13, and    arrow-shaped prime market stage leaves, as in hybrid spinach variety    51-330, representative seed of which have been deposited under NCIMB    Accession NCIMB 41821, and is modified in one or more other    characteristics.-   16. Progeny as paragraph 15, wherein the modification is effected by    mutagenesis.-   17. Progeny as paragraph 15, wherein the modification is effected by    transformation with a transgene.-   18. A method of producing an inbred spinach plant derived from    hybrid spinach variety 51-330, comprising the steps:    -   a. preparing a progeny plant derived from hybrid spinach variety        51-330 by crossing the plant of paragraph 1 with a second        spinach plant;    -   b. crossing the progeny plant with itself or a second spinach        plant to produce a seed of a progeny plant of a subsequent        generation;    -   c. growing a progeny plant of a subsequent generation from said        seed and crossing the progeny plant of a subsequent generation        with itself or a second spinach plant; and    -   d. repeating step b) or c) for at least 1 more generation to        produce an inbred spinach plant derived from the hybrid spinach        variety 51-330.-   19. An inbred spinach plant produced by the method of paragraph 21.-   20. A method for producing spinach leaves as a fresh vegetable    comprising packaging leaves of a plant of paragraph 1 or paragraph 2    or paragraph 3 or paragraph 4 or paragraph 5.-   21. A method for producing spinach leaves as a processed food    comprising processing leaves of a plant of paragraph 1 or paragraph    2 or paragraph 3 or paragraph 4 or paragraph 5.-   22. One or more spinach plants of paragraph 1 or paragraph 2 or    paragraph 3 or paragraph 4 or paragraph 5, in a container, for    harvest of leaves.-   23. A method of determining the genotype of a plant of hybrid    spinach variety 51-330, representative seed of which having been    deposited under NCIMB Accession NCIMB 41821, or a first generation    progeny thereof, comprising obtaining a sample of nucleic acids from    said plant and detecting in said nucleic acids a plurality of    polymorphisms, wherein the plurality of polymorphisms are indicative    of and/or give rise to the expression of the morphological and    physiological characteristics of inbred hybrid spinach variety    51-330.-   24. The method of paragraph 23 additionally comprising the step of    storing the results of detecting the plurality of polymorphisms on a    computer readable medium, or transmitting the results of detecting    the plurality of polymorphisms.-   25. The computer readable medium of paragraph 24.

Having thus described in detail preferred embodiments of the presentinvention, it is to be understood that the invention is not to belimited to particular details set forth in the above description as manyapparent variations thereof are possible without departing from thespirit or scope of the present invention.

What is claimed is:
 1. A hybrid spinach variety designated 51-330exhibiting a combination of traits including resistance against downymildew (Peronospora farinosa f. sp. spinaciae) strains Pf1-Pf13, andarrow-shaped prime market stage leaves, representative seed of whichhaving been deposited under NCIMB Accession
 41821. 2. A seed of theplant of claim 1, wherein a plant grown from said seed has all themorphological and physiological characteristics of hybrid spinachvariety designated 51-330.
 3. A part of the plant of claim 1, whereinsaid part of the plant is suitable for vegetative reproduction whereinsaid part is a cutting, a root, a stem, a cell or a protoplast.
 4. Atissue culture of regenerable cells or protoplasts from the spinachplant of claim 1, wherein said cells or protoplasts of the tissueculture are derived from a tissue comprising a leaf, an embryo, acotyledon, a hypocotyl, a meristematic cell, a root, a root tip, ananther, a flower or a stem.
 5. A method of producing an inbred spinachplant derived from hybrid spinach variety 51-330, representative seed ofwhich having been deposited under NCIMB Accession 41821, comprising thesteps: a. preparing a progeny plant derived from hybrid spinach variety51-330 by crossing the plant of claim 4 with a second spinach plant; b.crossing the progeny plant with itself or a second spinach plant toproduce a seed of a progeny plant of a subsequent generation; c. growinga progeny plant of a subsequent generation from said seed and crossingthe progeny plant of a subsequent generation with itself or a secondspinach plant; and d. repeating step b) and c) for at least 1 moregeneration to produce an inbred spinach plant derived from the hybridspinach variety 51-330.
 6. A method for producing spinach leaves as afresh vegetable comprising packaging leaves of a plant of claim
 1. 7. Amethod for producing spinach leaves as a processed food comprisingprocessing leaves of a plant of claim
 1. 8. A container comprising oneor more spinach plants of claim 1 for harvest of leaves.
 9. A method ofdetermining the genotype of a plant of hybrid spinach variety 51-330,representative seed of which having been deposited under NCIMB Accession41821, comprising obtaining a sample of nucleic acids from said plantand detecting in said nucleic acids a plurality of polymorphisms,wherein the plurality of polymorphisms are indicative of and/or giverise to the expression of the morphological and physiologicalcharacteristics of inbred hybrid spinach variety 51-330.
 10. The methodof claim 9 additionally comprising the step of storing the results ofdetecting the plurality of polymorphisms on a computer readable medium,or transmitting the results of detecting the plurality of polymorphisms.11. The hybrid spinach variety of claim 1, which is a plant grown fromseed having been deposited under NCIMB Accession
 41821. 12. A spinachplant having all the morphological and physiological characteristics ofthe plant as claimed in claim 1, representative seed of which havingbeen deposited under NCIMB Accession No.
 41821. 13. A spinach plantproduced by transforming the spinach plant of claim 4 with a transgene.14. A seed capable of growing into a plant of the hybrid spinach varietyof claim 1.